Title

Authors

Published in:

Biophysical Journal 99,12 (2010) 3880-3886;

Link to original published article:

http://dx.doi.org/10.1016/j.bpj.2010.10.029

Abstract

Diffusion of two Escherichia coil outer membrane proteins-the cobalamin (vitamin B12) receptor (BtuB) and the OmpF porin, which are implicated in the cellular import pathways of colicins and phages-was measured in vivo. The lateral mobility of these proteins is relevant to the mechanism of formation of the translocon for cellular import of colicins such as the rRNase colicin E3. The diffusion coefficient (D) of BtuB, the primary colicin receptor, complexed to fluorescent antibody or colicin, is 0.05 +/- 0.01 mu m(2)/s and 0.10 +/- 0.02 mu m(2)/s, respectively, over a timescale of 25-150 ms. Mutagenesis of the BtuB TonB box, which eliminates or significantly weakens the interaction between BtuB and the TonB energy-transducing protein that is anchored in the cytoplasmic membrane, resulted in a fivefold larger value of D, 0.27 +/- 0.06 mu m(2)/S for antibody-labeled BtuB, indicating a cytoskeletal-like interaction of TonB with BtuB. OmpF has a diffusion coefficient of 0.006 +/- 0.002 mu m(2)/s, similar to 10-fold smaller than that of BtuB, and is restricted within a domain of diameter 100 nnn, showing it to be relatively immobile compared to BtuB. Thus, formation of the outer membrane translocon for cellular import of the nuclease colicins is a demonstrably dynamic process, because it depends on lateral diffusion of BtuB and collisional interaction with relatively immobile OmpF.